LOCUS       sequence-180126-        4407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of shRNA targeting CHRNA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4407)
  TITLE     Modiano Lab plasmids
REFERENCE   2  (bases 1 to 4407)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: alpha3-nAChR shRNA (Alpha-3.29)
FEATURES             Location/Qualifiers
     source          1..4407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..215
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     136..155
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(288..304)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(289..305)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(333..351)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(372..388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(372..388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(385..407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    396..412
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(420..426,427..444,445..450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    465..486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(603..620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(774..1362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(854..873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1533..2324,2325..2393))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2156..2175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2394..2498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2524..2979)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2661..2682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2873..2892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3106..3128
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3120..3137
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3121..3137
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3144..3163
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3144..3162
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3195..3211
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3195..3211
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(3274..3873)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3357..3377)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3854..3873
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3898..4397)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3946..3965)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     4339..4362
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      181 gatttgggaa tcttataagt tctgtatgag accacagatc ccaagtgtca agtatattgc
      241 tgttcaagag atcagcaata tacttgacat tttttggaaa agcttatcga taccgtcgac
      301 ctcgaggggg ggcccggtac cagcttttgt tccctttagt gagggttaat ttcgagcttg
      361 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      421 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      481 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      541 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      601 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1201 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1321 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1381 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1441 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1501 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1561 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1621 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1681 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1741 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1801 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1861 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1921 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1981 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2041 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2101 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2161 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2221 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2281 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2341 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2401 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2461 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2521 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2581 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2641 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2701 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2761 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2821 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2881 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2941 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     3001 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     3061 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     3121 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gagctccacc
     3181 gcggtggcgg ccgctctaga actagtggat cccccgggct gcatggggtc gtgcgctcct
     3241 ttcggtcggg cgctgcgggt cgtggggcgg gcgtcaggca ccgggcttgc gggtcatgca
     3301 ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc cgagccgctc
     3361 gtagaagggg aggttgcggg gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc
     3421 ggccgcctcc actccgggga gcacgacggc gctgcccaga cccttgccct ggtggtcggg
     3481 cgagacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg gcgccaggag
     3541 gccttccatc tgttgctgcg cggccagccg ggaaccgctc aactcggcca tgcgcgggcc
     3601 gatctcggcg aacaccgccc ccgcttcgac gctctccggc gtggtccaga ccgccaccgc
     3661 ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag
     3721 ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc
     3781 ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt
     3841 ggcgaggcgc accgtgggct tgtactcggt catggtaagc tagcttgggc tgcaggtcga
     3901 aaggcccgga gatgaggaag aggagaacag cgcggcagac gtgcgctttt gaagcgtgca
     3961 gaatgccggg cctccggagg accttcgggc gcccgccccg cccctgagcc cgcccctgag
     4021 cccgcccccg gacccacccc ttcccagcct ctgagcccag aaagcgaagg agcaaagctg
     4081 ctattggccg ctgccccaaa ggcctacccg cttccattgc tcagcggtgc tgtccatctg
     4141 cacgagacta gtgagacgtg ctacttccat ttgtcacgtc ctgcacgacg cgagctgcgg
     4201 ggcggggggg aacttcctga ctaggggagg agtagaaggt ggcgcgaagg ggccaccaaa
     4261 gaacggagcc ggttggcgcc taccggtgga tgtggaatgt gtgcgaggcc agaggccact
     4321 tgtgtagcgc caagtgccca gcggggctgc taaagcgcat gctccagact gccttgggaa
     4381 aagcgcctcc cctacccggt agaattc
//
